钙敏感受体及钙蛋白酶在大鼠脊髓神经元缺氧-复氧损伤中的表达及其意义  

Alterated expression and role of calcium sensing receptor and calpain induced by anoxia / reoxygenation in rat spinal cord neurons

作　　者：蒋润宇 孙继芾[1] 何炜[1] 张志坚[2] 黄永辉[1] 

机构地区：[1]江苏大学附属医院骨科,江苏镇江212001 [2]江苏大学医学院基础与临床医学研究中心,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2016年第3期208-211,214,共5页Journal of Jiangsu University：Medicine Edition

摘　　要：目的:探究大鼠脊髓神经元缺氧-复氧损伤中钙敏感受体及钙蛋白酶的表达变化及其意义。方法:将原代培养的新生SD大鼠脊髓神经元随机分成4组:正常对照组、缺氧-复氧组、激活组、抑制组。后3组神经元均制备缺氧-复氧损伤模型,激活组加入钙敏感受体激动剂Gd Cl3,抑制组加入受体抑制剂NPS2390。应用Hoechst 33258染色观察各组脊髓神经元凋亡;应用免疫荧光技术分析钙敏感受体及钙蛋白酶蛋白在脊髓神经元的表达定位;采用蛋白质印迹法检测各组钙敏感受体、钙蛋白酶以及Bax蛋白的表达水平。结果:与对照组比较,缺氧-复氧组脊髓神经元凋亡增加,并且钙敏感受体、钙蛋白酶、Bax蛋白的表达明显升高(均P<0.01);Gd Cl3可明显增强上述作用,而NPS2390可明显抑制上述作用。结论:在缺氧-复氧损伤过程中,脊髓神经元凋亡增加,钙敏感受体及钙蛋白酶表达增多。Objective： To investigate the aherated expression and role of calcium sensing receptor （CaSR） and calpain in apoptosis of rat spinal cord neurons induced by anoxia/reoxygenation （A/R）. Methods： After spinal anoxia/reoxygenation model was established by culturing primary spinal cell of neo- natal rat in vitro, the spinal cells were divided into four groups randomly：the control group, A/R group, cal- cimimetic group （ GdC13 group） and inhibitor group （ NPS2390 ）. The apoptotic rate of spinal cells was deter- mined by Hoechst 33258 assay, the expression of CaSR and calpain was detected by fluorescence immunoas- say and Western blotting. Results： In A/R group, there was a significant increase in the apoptotic rate and the levels of CaSR, calpain and Bax compared to the control group（P 〈0.01 ）. Howerer,the above markers increased more in GdCI3 group, and these parameters decreased in inhibitor group. Conclusion： During the process of hypoxia/reoxygenation injury in rat spinal cord neurons, the expressions CaSR and calpain in- creased, accompanied with increasing of the apoptosis rate.

关 键 词：钙敏感受体 钙蛋白酶 脊髓神经元 缺氧-复氧 凋亡 

分 类 号：R364.1[医药卫生—病理学]